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Journal of

Clinical Medicine

Article
The Impact of Virtual Reality Training on the Quality
of Real Antromastoidectomy Performance
Wojciech Gaw˛ecki 1, * , Magdalena W˛egrzyniak 1 , Patrycja Mickiewicz 2 ,
Maria Bratumiła Gawłowska 3,4 , Marcin Talar 3,5 and Małgorzata Wierzbicka 1
1 Department of Otolaryngology and Laryngological Oncology, Poznań University of Medical Sciences,
60-355 Poznań, Poland; m.wegrzyniak89@gmail.com (M.W.); otosk2@gmail.com (M.W.)
2 WSB University, 41-300 Dabrowa
˛ Górnicza, Poland; p.mickiewicz86@wp.pl
3 Medicus sp. z o.o., 50-224 Wrocław, Poland; mgawlowska@medicus.com.pl (M.B.G.);
mtalar@medicus.com.pl (M.T.)
4 Poznań University of Medical Sciences, 61-701 Poznań, Poland
5 Faculty of Health Sciences, Pomeranian Medical University in Szczecin, 71-210 Szczecin, Poland
* Correspondence: wojgaw@interia.pl; Tel.: +48-61-8691-387; Fax: +48-61-8691-690

Received: 10 September 2020; Accepted: 29 September 2020; Published: 2 October 2020 

Abstract: Background: The aim of this paper is to analyze the results of virtual reality (VR)
antromastoidectomy simulation training and the transferability of the obtained skills to real temporal
bone surgery. Methods: The study was conducted prospectively on a group of 10 physicians, and was
composed of five VR simulation training sessions followed by live temporal bone surgery. The quality
of performance was evaluated with a Task-Based Checklist (TBC) prepared by John Hopkins Hospital.
Additionally, during every VR session, the number and type of mistakes (complications) were noted.
Results: The quality of performance measured by the TBC increased significantly during consecutive
VR sessions. The mean scores for the first and fifth sessions were 1.84 and 4.27, respectively (p < 0.001).
Furthermore, the number of mistakes in consecutive VR sessions was gradually reduced from
11 to 0. During supervised surgery, all the participants were able to perform at least part of an
antromastoidectomy, and the mean TBC score was 3.57. There was a significant strong positive
correlation between the individual results of the fifth VR session and the individual results of
supervised surgery in the operating room (rp = 0.89, p = 0.001). Conclusions: Virtual reality for
temporal bone training makes it possible to acquire surgical skills in a safe environment before
performing supervised surgery. Furthermore, the individual final score of virtual antromastoidectomy
training allows a prediction of the quality of performance in real surgery.

Keywords: virtual reality; temporal bone surgery; antromastoidectomy; active assisting;


supervised surgery

1. Introduction
To date, temporal bone training has been largely based on traditional dissection of cadaveric
temporal bones through in-house training or participation in national or international temporal bone
courses [1]. Training facilities are in the vast majority of European ENT Departments dealing with
otosurgery, but due to the poor availability of human temporal bones, access for trainees is limited [1–3].
Virtual reality (VR) simulation and artificial temporal bone models have been gaining popularity as
training supplements to “wet” dissection in many institutions, and nowadays, multiple VR simulators
are commercially available [4–8].
VR training has many important advantages. It provides a wide range of anatomic variants
and enables failures without consequences and unlimited practice [9–13]. The key components of

J. Clin. Med. 2020, 9, 3197; doi:10.3390/jcm9103197 www.mdpi.com/journal/jcm


J. Clin. Med. 2020, 9, 3197 2 of 11

the training are to avoid exceeding the anatomical boundaries of a complete mastoidectomy and to
avoid violating vital structures [6]. Simulation-based technical skills training must present a good
balance between automatic scoring, drilling time and self-assessment [14]. A simulator-integrated
tutor function has significantly improved performance in these areas [6], and the number of objective
assessment tools for self-directed practice has been reported [15]. A feasible alternative is final-product
analysis or a simulator-based automated assessment and feedback scoring system [14,15]. For a
combined metrics-based score (MBS), a significant discriminative ability between experienced
surgeons and residents was demonstrated; nevertheless, it failed to measure or encourage safe
routines [14]. Moreover, at present, the automated feedback based on metrics in VR simulation
does not have a sufficient empirical basis and has not been generally accepted for use in training
or certification [16]. Thus, the VR simulation training requires supplemental approaches, feedback,
improved self-assessment tools and raising awareness of real operating field challenges. Nearly all
previous VR studies were limited because that they did not assess the effect of simulation training on
“real life” performance in an operating room (OR).
Thus, the aim of this paper is to analyze the results of VR simulation training for
antromastoidectomy, which is one of the basic otosurgical procedures [17], and to evaluate the
transferability of the obtained skills to the real temporal bone operating field, measured by a specialist’s
assessment of a supervised surgery.

2. Experimental Section

2.1. Study Design and Setting


The study was conducted prospectively, with five VR-simulation training sessions (separated
by 4–5-week breaks), arranged before the real temporal bone surgery in the OR (Figure 1).
Participants performed a series of virtual dissections after the preacquaintance and demonstration
session with the VR system. During every VR session, the participants had to perform a virtual
antromastoidectomy (cortical mastoidectomy). After the completed VR training, their knowledge was
checked while actively assisting in a mastoid process surgery. The participants’ performance was
then scored during a supervised surgery—an antromastoidectomy performed as a part of a cochlear
implantation. Participants were allowed to assist or perform any real surgical procedures only if they
were able to perform the fifth VR simulation without any mistakes (complications). The study was
performed between October 2018 and August 2019, and the regional ethics committee deemed this
study to be exempt.

2.2. Materials
The participants consisted of 10 physicians: four ENT specialists experienced in rhinology or head
and neck oncology, and six otorhinolaryngology residents. None of the participants were experienced
in otosurgery. They had never performed an antromastoidectomy before, neither in a VR simulator
or cadaver specimen nor in a real patient in an OR. All of them had assisted in many (more than
30) otological procedures (tympanoplasties, cholesteatoma surgeries and cochlear implantations).
Two experienced otosurgeons (with more than 10 years of experience and more than 1000 performed
middle-ear surgeries) participated in the study as supervisors for all the participants.
J. Clin. Med. 2020, 9, 3197 3 of 11
J. Clin. Med. 2020, 9, x FOR PEER REVIEW 3 of 11

PRE-ACQUAINTANCE AND DEMONSTRATION SESSION


WITH THE VR SYSTEM

VR SESSION 1 - PERFORMANCE AND EVALUATION

VR SESSION 2 - PERFORMANCE AND EVALUATION

VR SESSION 3 - PERFORMANCE AND EVALUATION

VR SESSION 4 - PERFORMANCE AND EVALUATION

VR SESSION 5 - PERFORMANCE AND EVALUATION


IF NO ERRORS / FAILURES

ACTIVE ASSISTING DURING SURGERY

SUPERVISED SURGERY
Figure 1. Study
Figure design.
1. Study design.

2.2. 2.3. VR Simulator


Materials
TheThe participants
participants were presented
consisted with a graphical
of 10 physicians: four ENT representation of the bone via
specialists experienced a temporalorbone
in rhinology
head surgery simulator
and neck composed
oncology, of aotorhinolaryngology
and six Geomagic Touch Haptic Device None
residents. from 3D of Systems, a MIDI controller
the participants were
from KORG,
experienced NVidia 3D They
in otosurgery. Active glasses,
had neveraperformed
monitor with an 3D technology, workstation
antromastoidectomy before,PC and software
neither in a
created by a team from The University of Melbourne. The system was validated
VR simulator or cadaver specimen nor in a real patient in an OR. All of them had assisted in many for its user interface
(moreandthan
content
30)[7,18,19]. Theprocedures
otological models were generated from microCT
(tympanoplasties, human surgeries
cholesteatoma temporal boneand scans
cochlearwith a
voxel resolution
implantations). Two of 96 × 96 × 96
experienced µm [20]. Thanks
otosurgeons to thethan
(with more 3D Active glasses,
10 years 3D monitor
of experience and andmorededicated
than
1000software,
performedthe system gives surgeries)
middle-ear access to a participated
three-dimensional
in theview.
studyIn this system, the drill
as supervisors handpiece
for all the
with a cutting or polishing tip is visible on the screen. The tips can be changed depending on the
participants.
needs. The drill handpiece and the tip accurately reflect what the surgeon can see in the real operating
2.3. field.
VR Simulator
The handle of the haptic device resembles the weight of a drill used in real surgical operations.
While working with
The participants the presented
were haptic device,
withthe user feels vibrations
a graphical whileofdrilling,
representation the bone resistance to stronger
via a temporal
bone pressure
surgeryon the hardcomposed
simulator bone structures in the 3DTouch
of a Geomagic modelHaptic
and additionally,
Device from hears the changing
3D Systems, a MIDIsound.
The remains of the drilled bone are invisible to the user. The model can
controller from KORG, NVidia 3D Active glasses, a monitor with 3D technology, workstation PC be rotated in any direction.
andIn our study,
software we selected
created onefrom
by a team simple (and
The always the
University of same) modelThe
Melbourne. of asystem
temporal bone
was with very
validated fortypical
its
anatomical features, which reflects most of the population’s temporal
user interface and content [7,18,19]. The models were generated from microCT human temporal bones. A study participant
bone during
scansawith
VR session
a voxelisresolution
presented ofin Figure
96 × 962.× 96 µm [20]. Thanks to the 3D Active glasses, 3D
monitor and dedicated software, the system gives access to a three-dimensional view. In this system,
the drill handpiece with a cutting or polishing tip is visible on the screen. The tips can be changed
depending on the needs. The drill handpiece and the tip accurately reflect what the surgeon can see
in the real operating field. The handle of the haptic device resembles the weight of a drill used in real
surgical operations. While working with the haptic device, the user feels vibrations while drilling,
J. Clin. Med. 2020, 9, 3197 4 of 11

Figure 2. Virtual reality (VR) temporal bone surgery.

2.4. Evaluation of VR Sessions


The quality of performance was evaluated by a Task-Based Checklist (TBC) prepared by John
Hopkins Hospital and described by Francis et al. [21]. The TBC is used to evaluate the performance
of a series of individual surgical steps on a 5-point Likert scale—from 1 (unable to perform) to 5
(performs easily with good flow). The whole checklist, prepared for evaluation of middle-ear surgery,
is composed of 22 items, grouped into seven major tasks. In our study, 11 items, grouped into
the following four tasks concerning cortical mastoidectomy were evaluated: (1) initial bone cuts,
(2) defining anatomic limits, (3) open antrum and (4) thin posterior EAC (external auditory canal)
cortex. Additionally, during every VR session, the number and type of mistakes (complications) were
noted. Every simulation was assessed by both supervisors, and a mean of two ratings was calculated.
The time of the session was limited to 40 min.

2.5. Active Assisting During Surgery


The participants who had no failures in the fifth VR simulation were allowed to actively assist
during a surgery. The surgery was always performed by the same expert supervising the study (WG).
During active assisting surgery, the participants were assessed on their task of verbally listing the
following surgical steps in the proper order: (1) removal of the cortical bone, (2) opening of the mastoid
air cells, (3) visualization of the dura and sigmoid sinus, (4) finding the antrum, (5) finding the lateral
semicircular canal and (6) finding the short process of the incus. Furthermore, they were asked to give
the proper names of the following anatomical details indicated by the otosurgeon: (1) the posterior
wall of the external auditory canal, (2) the middle cranial fossa dura, (3) the sigmoid sinus, (4) the
lateral semicircular canal, (5) the incus and (6) the course of the facial nerve.

2.6. Supervised Surgery


Supervised surgery was always performed under the guidance of the same supervising otosurgeon
(WG), and was always evaluated by both supervising experts. Participants had to perform an
antromastoidectomy as part of a cochlear implantation procedure. The procedures were performed
only on patients with normal anatomy of the temporal bone, as confirmed by a preoperative CT.
The quality of performance was evaluated by the TBC in the same way as the VR training sessions.
During real surgery, the safety of the patient was crucial and no mistakes were allowed, thus the
J. Clin. Med. 2020, 9, 3197 5 of 11

surgery would be stopped (not completed by the participant) if a supervisor decided it was potentially
dangerous or the participant did not feel confident enough to continue. The time limit was 40 min.

2.7. Statistical Analysis


Statistical analysis was carried out in R software, version 3.5.1, 2018 (http://cran.r-project.org),
The R Foundation for Statistical Computing Institute for Statistics and Mathematics, Vienna, Austria.
Normality of TBC data was confirmed based on Shapiro–Wilk test results, as well as based on skewness
and kurtosis values and visual assessment of histograms. TBC results between sessions were compared
with MANOVA, a Mauchly test was used to check data sphericity, and Greenhouse–Geisser and
Huynh–Feldt corrections for departure from sphericity were applied. To identify specific sessions
that had a significantly different TBC result, a post-hoc test was used (a paired t-test with Bonferroni
correction for multiple comparisons). Correlation between the last VR session and the OR session for
TBC was analyzed with Pearson’s correlation coefficient (rp ). The intraclass correlation coefficient
(ICC) was computed to assess the agreement between the two raters. All tests were based on α = 0.05.

3. Results
The group of 10 participants managed to complete the course of five VR sessions, and all were
allowed to take part in active assisting during a surgery, and further, in a supervised surgery.

3.1. The Quality of the Performance of VR Simulation


The quality of performance measured by TBC increased significantly during consecutive VR
sessions. The mean scores for the first and fifth sessions were 1.84 and 4.27, respectively (p < 0.001).
There was a significant difference between all five VR sessions in the average TBC result, as based on
the MANOVA analysis (p < 0.001). To identify which sessions were responsible for this difference,
a post-hoc test was conducted. The results are presented in Tables 1 and 2, and in Figure 3.

Figure 3. Boxplot of TBC results between sessions (the circle above VR2 represents an outlier
observation).
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Table 1. Task-Based Checklist (TBC) results by session (VR–virtual reality surgery; OR–supervised
surgery in operating room; Q1–lower quartile; Q3–upper quartile; n = 10).

Session Mean SD Median Q1 Q3 Min. Max.


VR1 1.84 0.41 1.86 1.56 2.08 1.18 2.50
VR2 2.41 0.68 2.43 2.20 2.80 1.18 3.32
VR3 3.10 0.64 3.18 3.02 3.60 2.00 3.77
VR4 3.70 0.76 4.09 3.31 4.23 2.32 4.41
VR5 4.27 0.64 4.61 3.74 4.76 3.27 4.95
OR 3.57 0.67 3.61 2.90 4.10 2.77 4.68

Table 2. TBC change between sessions (MANOVA post-hoc test; MD—mean difference in TBC
between two sessions calculated as the later session mean minus the previous session mean, with a
95% confidence interval (CI); p–individual paired t-test p-value; padj –paired t-test p-value adjusted for
multiple comparisons (Bonferroni correction)).

Sessions Comparison MD 95% CI p padj


VR1–VR2 0.57 (0.32; 0.82) 0.001 0.009
VR1–VR3 1.26 (0.96; 1.56) <0.001 <0.001
VR1–VR4 1.85 (1.47; 2.24) <0.001 <0.001
VR1–VR5 2.43 (2.07; 2.78) <0.001 <0.001
VR1–OR 1.73 (1.27; 2.19) <0.001 <0.001
VR2–VR3 0.69 (0.53; 0.84) <0.001 <0.001
VR2–VR4 1.28 (0.99; 1.57) <0.001 <0.001
VR2–VR5 1.85 (1.49; 2.22) <0.001 <0.001
VR2–OR 1.16 (0.64; 1.68) 0.001 0.010
VR3–VR4 0.60 (0.40; 0.79) <0.001 0.001
VR3–VR5 1.17 (0.90; 1.44) <0.001 <0.001
VR3–OR 0.47 (0.04; 0.91) 0.036 0.534
VR4–VR5 0.57 (0.40; 0.74) <0.001 <0.001
VR4–OR −0.12 (−0.49; −0.24) 0.467 >0.999
VR5–OR −0.70 (−0.92;−0.47) <0.001 0.001

3.2. The Number of Mistakes in VR Simulations


In consecutive VR sessions, the number of mistakes was evidently reduced. Altogether, 11 mistakes
were observed in the participants’ first session (among them, two injuries of the facial nerve and
four injuries of the inner ear). No mistakes were observed in their fifth session. The number and
characteristics of the failures are presented in Figure 4.

3.3. Evaluation of Active Assisting During Surgery


Due to the fulfillment of the threshold criterion of no mistake during the final VR session, all the
participants were allowed to take part in active assisting during a surgery. During it, all the participants
were able to list the surgical steps in the proper order, and all but one were able to give the proper
names of all the indicated anatomical details. Only one resident had a problem giving the proper name
of the incus, but named the other structures properly.
J. Clin. Med. 2020, 9, 3197 7 of 11

12

10

8
Number of failures

damage to the middle cranial


fossa dura
damage to the sigmoid sinus
2
damage to the incus (visible
erosion)
damage to the posterior wall
of the external auditory canal
0 damage to the inner ear (hole
0 1 2 3 4 5 to the labirynth)
Session damage to the facial nerve

Figure 4. The number and characteristics of the failures during VR sessions.

3.4. Evaluation of a Supervised Antromastoidectomy Performance


All of the participants of the study were then allowed to perform a supervised surgery. The mean
score was 3.57, which was better than for the first, second and third VR sessions (the difference for
Sessions 1 and 2 was statistically significant), although worse than the fourth and fifth VR sessions
(for Session 5, the difference was statistically significant). The results are presented in Tables 1 and 2,
and in Figure 3. Five of the ten supervised surgeries were not completed by the participant, requiring
the supervisor to finish. Reasons included: (1) participant’s decision (lack of self-confidence to
continue) in two cases, (2) supervisor’s decision (surgery being potentially dangerous) in one case and
(3) exceeding the time limit in two cases.

3.5. Correlation of the VR Final Score and Real Antromastoidectomy Performance


There was a significant strong positive correlation between the individual results of the fifth VR
session and the individual results of supervised surgery in the OR (rp = 0.89, p = 0.001). The details are
presented in Figure 5.

3.6. Inter-Rater Agreement


There was significant excellent agreement between raters for all the sessions, using the two-way
random effect models, ICC = 0.995; CI95 [0.981; 0.998]; p < 0.001.
J. Clin. Med. 2020, 9, 3197 8 of 11

Figure 5. Scatterplot for TBC result between VR5 session and operating room (OR) surgery.

4. Discussion
The aim of this paper was to analyze the results of VR simulation training with special regard to the
transferability of the obtained surgical skills to the real temporal bone operating field. The influence of
VR training on different real surgeries, i.e., colonoscopy, laparoscopic camera navigation and endoscopic
sinus surgery, showed that the participants who reached proficiency in simulation-based training
performed better in a patient-based setting than their counterparts who did not have such training,
and furthermore, simulation-based training was equally as effective as patient-based training [22].
In temporal bone surgery, human cadaveric dissection has been the gold standard of training for a
long time because it closely mimics real-life surgical conditions [23]. However, contemporary otologic
training is primarily acquired in an OR because the instruction and practice of using a cadaveric
temporal bone is less consistently available to trainees today [11]. Furthermore, artificial bone models,
which are popular in multiple medical disciplines (dental implants, maxillo-facial surgery, orthopedics),
are not widely used in otosurgery because of some deficiencies in comparison to real temporal bone.
However, they can be an alternative for beginners [24,25].
Fortunately, a new option—VR temporal bone training—has been gaining importance for the
past few years, and nowadays, VR is a well-established and useful adjunct to traditional cadaveric
dissection of temporal bone for trainees [6,19,26]. Such training may play an important and increasing
role in education in the future, but under certain conditions and according to some standards [14].
This way of training is reportedly used in many leading training departments in Europe, and most of
the remaining departments expect to implement VR simulation for temporal bone training into their
residency programs in the near future [1].
J. Clin. Med. 2020, 9, 3197 9 of 11

The question arises whether this kind of training would allow participants to skip ahead,
from VR to gradually more advanced assists and supervised surgeries, and consequently to
replacing cadaveric temporal bone dissections completely or significantly. The benefits of VR
training in antromastoidectomy performance were shown in cadaveric temporal bone dissection [19].
Andersen et al. proved that even two hours of self-directed VR simulation training were effective in
increasing cadaveric dissection mastoidectomy performance. The conclusion is that mastoidectomy
skills are transferable from VR simulation to traditional dissection [6]. The impact of VR training on real
middle-ear surgery performance was checked by Al-Noury, who found that in previously simulated
cases, the residents scored higher, were faster and more confident and required fewer instructions [26].
However, the impact of VR training on real surgery has not been sufficiently proved. Thus, in this
paper, we aimed at answering whether VR is an effective training instrument that enables physicians to
obtain skills sufficient for surgery in a real temporal bone operating field. For this purpose, we planned
a training program of five sessions separated by 4–5-week breaks. This was based on the observations
described by Andersen et al., who found that the mastoidectomy skills acquired under time-distributed
practice conditions were retained better than skills acquired under massed practice conditions [27].
The tool we used was a validated Task-Based Checklist (TBC), prepared by John Hopkins
Hospital [21]. According to Sethia et al., who reviewed the literature for assessment of performance for
mastoidectomy, this tool possesses the most validity evidence of those reviewed [15]. However, as TBC
does not take into account the number and type of performed mistakes (intraoperative complications)
during a surgery, we added such analysis in every VR session. We found that the VR lab is perfect
to facilitate repeated training in mastoid basic surgery. During consecutive sessions, the quality of
performance increased, and most importantly, the number of mistakes (intraoperative complications)
gradually decreased to zero. This effect can be explained by both an improvement in anatomical
knowledge and surgical skills after repeated training. Active assisting during surgery confirmed
excellent anatomical and surgical knowledge obtained during the VR antromastoidectomy course.
All of the participants were then allowed to perform a supervised surgery, and all of them managed to
perform at least part of an antromastoidectomy, with a mean score that compared to that between their
third and fourth VR sessions. Such results indicate the importance of VR training (results significantly
better than in the first and second VR sessions), and also the influence of the change of working
environment from VR to real surgery (where results were significantly worse in comparison to the fifth,
and last, VR session), which is fully understandable. As safety during the surgery and the wellbeing of
the patient are always the most important aspect, it is understandable that either the participant or
supervisor decided to stop when the safety of the surrounding structures was potentially endangered
or if the surgery time was exceeded. No complications occurred during the surgeries performed by
the participants or in combination with the supervisors. We have shown that just five sessions of VR
temporal bone training might be enough to enable physicians to obtain the skills sufficient to perform
a supervised antromastoidectomy. Furthermore, we found a significant strong positive correlation
between the individual results of the fifth VR session and the individual results of the supervised
surgery in the OR, which shows that the individual final score of VR training allows a prediction of the
quality of performance in a real surgery.
This study has several limitations. First, the VR simulator used in the study was relatively simple.
We are aware that making VR simulations more realistic could improve the quality of surgery in the
operating room. Second, the lack of training on temporal bones from cadavers after the last VR session
and immediate switch to a real operation is a downside of this study, but at the same time, we are trying
to prove the possibility of bypassing that intermediate training stage, as it is increasingly difficult to
organize. The most serious study limitation is its small sample size and single-center design.
This study also has several advantages. The most noteworthy is the repeated use of the same VR
model and comparing the learning curve and gained skills with the in vivo operating field.
J. Clin. Med. 2020, 9, 3197 10 of 11

To summarize, VR temporal bone training can address needs in continuing education and
competency-based residency training, but there is still the open question of whether it ultimately
becomes a component of the certification process. We believe that in the future, this form of
repeated VR simulation can even totally replace the cadaveric temporal bone lab and allow residents
to proceed to surgery under specialist guidance, preselected by active assisting during a surgery.
Systematic integration of training using VR simulation is possibly the future direction of surgical
first steps.

5. Conclusions
Virtual reality for temporal bone training makes it possible to acquire surgical skills in a safe
environment before performing supervised surgery. Furthermore, the individual final score of virtual
antromastoidectomy training allows a prediction of the quality of performance in a real surgery.

Author Contributions: Conceptualization, M.W. (Małgorzata Wierzbicka) and W.G.; methodology, W.G., P.M.
and M.W. (Magdalena W˛egrzyniak); software, M.T.; formal analysis, W.G. and M.B.G.; investigation, W.G.
and M.W. (Magdalena W˛egrzyniak); resources, M.T.; data curation, W.G. and M.W. (Magdalena W˛egrzyniak);
writing, W.G. and M.W. (Małgorzata Wierzbicka); writing—review and editing, M.W. (Małgorzata Wierzbicka),
P.M. and W.G.; visualization, M.B.G.; supervision, M.W. (Małgorzata Wierzbicka); project administration, M.W.
(Małgorzata Wierzbicka). All authors have read and agreed to the published version of the manuscript.
Funding: This research received no external funding.
Acknowledgments: Authors wish to thank the Cochlear Ltd., Sydney, for free sharing of the VR equipment
whereby the study could be performed.
Conflicts of Interest: The authors declare no conflict of interest.

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